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Study on Effect of Pancreatic Juice Regurgitation on Acute Cholecystitis in Rabbit

Author: SongJianQiang
Tutor: FengZiTan
School: Hebei Medical University
Course: Internal Medicine
Keywords: Acute cholecystitis Common bile duct obstruction Pancreatic juice reflux Macrophages Tumor necrosis factor -α
CLC: R657.4
Type: Master's thesis
Year: 2008
Downloads: 35
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Abstract

Purpose: pancreatic duct the ampullary obstruction can cause cholestasis and pancreatic juice reflux. Cholestasis, pancreatic juice reflux can cause acute cholecystitis. Pancreatic juice reflux role in the pathogenesis of acute cholecystitis, not yet reported in the literature. The purpose of this test is by using rabbit bile duct ligation and simultaneous injection of trypsin to establish an animal model of acute cholecystitis, observe the common bile duct obstruction and bile amylase activity trypsin injection group and pure common bile duct obstruction group, serum the gallbladder tissue tumor necrosis factor-a, the gallbladder tissue macrophages quantity, gallbladder gross morphology different, to explore the mechanism of pancreatic juice regurgitation in acute cholecystitis; caused by stones and other factors for clinical pancreatic duct ampulla obstruction leads pancreatic juice reflux caused cholecystitis the basis of animal experiments, and also to provide new ideas for the future study of trypsin cholecystitis experimental, and a better understanding of the pathogenesis of acute cholecystitis. Methods: 45 healthy rabbits weighing 2.0-3.0kg / male or female, were randomly divided into sham operation group (SO), common bile duct obstruction group (BDL group), common bile duct obstruction and trypsin injection group ( group M), 15 each in 3 days killed, all rabbits before surgery day fasting, can not help but water. Obstruction of the common bile duct and pancreatic enzyme the injected group model preparation: ear vein injection of anesthesia with 3% sodium pentobarbital (30-35mg/kg) in aseptic downlink epigastric middle of the incision, blunt dissection of the common bile duct, duodenal opening of ligation of the common bile duct near first sterile Children vein catheter puncture above the ligation of the common bile duct, with silk above mild ligation puncture point (for the degree of non-bile leakage) 1ml no bacteria syringe leads to 0.2ml bile, extracted with a micropipette gun a 50U/ml trypsin 0.2ml injected into a 1 ml syringe, and injected into the common bile duct to 0.1ml/min extracted puncture tube, above the ligature. Checked blood and bile leakage after the abdomen was closed. Preparation of animal model of common bile duct obstruction groups: under the same conditions of anesthesia, laparotomy, blunt dissection of common bile duct, after the common bile duct near the duodenum openings double ligation. Sham group only blunt dissection after anesthesia and sterile laparotomy the common bile duct ligation. Modeling and placed in the cages were observed in rabbits in general, were killed three days later. Blood samples were drawn before death, centrifugal, detect serum levels of tumor necrosis factor-a; extract the gallbladder bile detect bile amylase; removal of the gallbladder, and take the gallbladder (ensure that the parts of each group taken), weighing 200mg homogenized After centrifugation, the supernatant was stored at -20 ℃ for radioimmunoassay of tumor necrosis factor-a, take the rest of the right amount gallbladder tissue in 10% formalin fixed, paraffin slice pathological examination and immunohistochemistry chemical methods to measure the number of macrophages in gallbladder tissue, the results of all the data were expressed as mean ± standard deviation among the groups were compared using one-way ANOVA, P lt; 0.05 was considered statistically significant. Results: 1 general form: visual observation of the the 1.1 common bile duct obstruction and trypsin injection group: rabbit gallbladder volume was significantly increased adhesion with the surrounding tissue and organs, the bottom surface of the gallbladder with large and small pus moss, dull color, partial mucosal necrosis small ulcers, gallbladder bile render the grass green, dark green, guts sludge formation; microscope: the mucosal epithelium severe off, even epithelial disappeared, vascular dilatation and congestion, mucosal lamina propria, muscularis, serosa high degree of edema were seen, a large number of inflammatory cell infiltration, muscularis vague, often accompanied by gallbladder inflammation around; 1.2 mere obstruction naked eye: gallbladder volume was increased, no adhesion with the surrounding organs, the appearance of pus point, mucosal integrity, wall edema, dark color, surface without ulcers, gallbladder bile render the grass green, dark green, biliary sludge formation; microscopy: shedding mild mucosal epithelium, lamina propria, muscularis, serosa has edema, vascular dilatation and congestion, and a small amount of inflammatory cell infiltration; 1.3 sham group naked eye: little gallbladder and surrounding organs, adhesions, look no pus points, mucosal integrity, slightly darker color, no ulcer surface, the gallbladder bile grass green, biliary sludge formation; microscope: mucosal epithelial intact layers edema, vascular congestion, and inflammatory cell infiltration. Sham operation group of 2 rabbits gallbladder bile starch enzyme activity (U / dL) changes gallbladder bile amylase activity (59.60 ± 8.77), simple common bile duct obstruction group gallbladder bile amylase activity (59.41 ± 9.20), common bile duct obstruction and trypsin injection group gallbladder bile amylase activity (166.08 ± 10.00); common bile duct obstruction and trypsin injection group gallbladder bile amylase activity significantly higher than the common bile duct obstruction alone group and the sham group (P lt; 0.05 ), and simple common bile duct obstruction group with the sham group no significant difference (P gt; 0.05). Changes of the value of tumor necrosis factor-a (× 10pg/ml) 3 rabbit serum and gallbladder tissue the sham group serum tumor necrosis factor-a (36.2 ± 4.49), and simple common bile duct obstruction serum levels of tumor necrosis factor -a (53.0 ± 6.41), common bile duct obstruction and trypsin the injected serum tumor necrosis factor-a (72.80 ± 6.47); common bile duct obstruction and trypsin injected serum tumor necrosis factor-a value significantly higher than those of bile Explorer obstruction group and the sham group (P lt; 0.05), simple common bile duct obstruction group was significantly higher than the sham group (P lt; 0.05). Sham group gallbladder tumor necrosis factor-a (7.66 ± 0.46), to simple common bile duct the obstruction group gallbladder tumor necrosis factor-a (26.30 ± 2.54), common bile duct obstruction and trypsin the injection group gallbladder tumor necrosis factor - a to (45.70 ± 2.79); common bile duct obstruction and trypsin the injected group gallbladder organization tumor necrosis factor-a value significantly higher than that of the common bile duct obstruction alone group and the sham group (P lt; 0.05), simple common bile duct obstruction group was significantly higher in the sham group (P lt; 0.05). 4 groups of rabbits gallbladder tissue changes of the number of macrophages in the sham group gallbladder tissue macrophages (4.33 ± 2.16), simple common bile duct obstruction groups gallbladder tissue macrophages (17.16 ± 3.86), bile Explorer obstruction and trypsin injection group gallbladder tissue macrophages (32.66 ± 6.91); common bile duct obstruction and trypsin injection group gallbladder tissue macrophages significantly higher than that of the common bile duct obstruction alone group and the sham group (P lt; 0.05), simple common bile duct obstruction was significantly higher than that in the sham group (P lt; 0.05). DAB stained dark brown macrophages, interstitial sham group rabbits gallbladder mucosa and submucosal tissue shows a small amount of macrophages, common bile duct obstruction alone group, common bile duct obstruction and trypsin injection group in the mucous membrane between quality and submucosal tissue macrophages increased significantly, and the common bile duct obstruction and the trypsin injection group was higher than that of common bile duct obstruction group increased significantly. Conclusions: 1 pancreatic juice reflux heavier biliary obstruction caused by acute cholecystitis, gallbladder pancreatic juice reflux gross morphology showed that inflammation is more pronounced. 2 pancreatic juice reflux gallbladder tissue macrophages increased further increase in serum and gallbladder tissue tumor necrosis factor-α, secretin regurgitation increased biliary obstruction caused by acute cholecystitis.

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